Orchid Flasking anyone?

[aNisip]

Does anybody dabble in the propogation of orchids? I have done A LOT of reading/researching but can't seem to find a true answer...maybe too many different sources...

I want to do the green pod method as all I have to steralize is the pod and tools...is it as simple as cutting open an immature seed pod after it soaked for 10min in bleach solution, then scooping out the seeds onto a general agar solution and then they'll grow? Some sources say I can use a petri dish while others say a flask is the only way...

Any info would be great!

All the best,

Andrew

 

[frogparty]

I am NOT a fan of the green pod method, nor do I like Petri dishes for any method of flasking. I have flanked several genera now, and always do my flasking at work in a bio safety cabinet, or laminar flow hood. Sterility is key. Get yourself a membership at www.orchidsinteractive.com and go through the flasking section. Great info to be had

 

[frogparty]

Petri dishes don't keep the humidity in like a flask does. You CAN use wide mouth mason jars...

 

[ScienceGirl]

Ooooh! Orchids! I've been reading about terrariums and how orchids love the humidity (provided they are large enough to give the plant room.) What do you guys think about keeping one in a terrarium? I was thinking about a cloche.

 

[aNisip]

Why do you dislike the greenpod method? From what I have read, the 'regular' way ends up with losing a lot of seeds bc the sterilization is too much for a lot of the vulnerable seeds...will like a general agar solution work?

 

[Tony C]

Flasking and tissue culture have been an idea I've tossed around for a while but just haven't had a chance to try. I would really like to experiment with Sarracenia TC, the process is still in its infancy and there are only a handful of clones in culture.

Ooooh! Orchids! I've been reading about terrariums and how orchids love the humidity (provided they are large enough to give the plant room.) What do you guys think about keeping one in a terrarium? I was thinking about a cloche.

Depends on the orchid, jewel types like Ludisia and Macodes are the most suitable for terrariums and many of the smaller Pleurothallids and Bulbophyllum group are also good candidates. Larger orchids like the common Phalaenopsis and Oncidium alliance intergenerics are generally too big, if the plant itself doesn't out grow the terrarium its inflorescence certainly will. Many are also prone to fungal and bacterial infection if kept in a location with poor airflow and high humidity.

 

[frogparty]

Andrew- with the green pod method, you risk losing just as many to being too immature to germinate. With good sterility, you don't lose too many, besides, out of the 1000s in a pod, how many do you want? One pod can make many flasks

Science girl- see what Vivarium orchids are all about at www.dendroboard.com

 

[angelofdeathzz]

There's no Orchids drinking out of a flask here! I kinda feel ripped off...

 

[frogparty]

Hahaha

 

[aNisip]

Hey frogparty so basically how should I go about doing this? (Phaelenopsis ap btw)

Wait til the seed pods brown and burst, then what?

Thanks,

Andrew

 

[frogparty]

AAAHHH- Heres the trick. Let them brown, but keep a close eye on them , and get them BEFORE they burst. I know its a pain, but you shoud start to see the ridges on the pod come under tension as the pod browns. What I like to do is get the pod off the inflorescense at that point, give it an EtOH spray down (70% is fine) and then, in a sterile draft free environment, open the pod yourself using sterile tools like a scalpel or even a razorblade along the ridges of the pod where they would naturally splt if allowed to dry a few more days. AS FOR WHAT MEDIA YOU CHOOSE TO USE, there are a myriad of options out there. Ill have to go through my in house lab notebook to find the one Ive had the most success with, BUT NOTE- I have never flasked Phalaenopsis seedlings, so results may differ, Id get on orchid board, or orchids interactive and see what people like best for flasking Phals. Get whatever flasks youre going to use and run them through the dishwasher, then fill 1/2 way with water and microwave 5 min on high. Put in your hood/glove box/ whatever and spray with 100% EtOH, and allow to thoroughly evaporate, pour your media in, allow to settle, cool, then alloquot out the contents of the pod into as many flasks as possible. I like wide mouth mason jars for this, because the seeds are easy to get in, and the seedlings are easy to get out!!!!!

http://www.orchidboard.com/community/propagation/9836-redneck-orchid-flasking.html

Redneck orchid flasking.....LOL! It really does work though

 

[frogparty]

ALSO- HUGE TIP!!! Go to www.fungi.com, and order their 0.2micron filter disks that fit on the jars!!!!!! Keep things sterile, but lets the flasks breathe, and keeps humidity high!!! MY SECRET WEAPON IN FLASKING SUCCESS

 

[aNisip]

Wow! Great info thanks a lot frogparty! Couple questions: can I run through dishwasher at home, plug them...then take to my lab at school where there is a hood where I can fill halfway microwave high then spray and let evaporate? And should I let it cool before I spray it? Or pick up with tongs and then spray it? And should I like rubberband the micron filters? Or how do I affix them?

And then take the brown un-opened pods and then open them manually and such and then sprinkle the seeds onto the medium in the mason jars, right? Or do I have to sterilize the seeds before I add to mason jars...

 

[frogparty]

the filter disks can be held on the jars by the ring part of the canning lid. these disks are DESIGNED to work with mason jars!

The dishwasher/microwave is assuming you dont have access to an autoclave- If you do, just foil the tops and autoclave your flasks at school. Once they have been run, place in the hood to cool. spray OUTSIDE of flasks with EtOH, and wipe down. Use gloves, spray your gloved hands. STERILE STERILE STERILE. The seeds inside the unopened pod are sterile. If you open the pod in the hood, youll be ok. THEN i WOULD USE AN INNOCULATING LOOP to remove seeds from pod and spread over media (cooled)

A lot of growers pour their flasks, and let the media set for a week to see if it contaminates before they innoculate. That way you dont waste good seeds on contaminated media.

Autoclaving your media for 60 min at 121C shoud do a good job of killing everything

 

[aNisip]

Would this be the right micron filters? http://www.fungi.com/product-detail/product/2-micron-filters-for-the-ealf47-set-of-10.html

 

[frogparty]

Those are the ones! Get the wide mouth jars so they fit properly

 

[aNisip]

Alright so basically:

1. Wait til they brown, not burst, collet them and store them in the dark.

2. Sterelize the pod with 70% EtOH

3. Get mason jars, fill half way with water and put them in dishwasher...then fill halfway with water in each and put each microwave on high for 5 min.

4. Then spray in and out with 100% EtOH and wait till it evaporates...

5. Then pour in agar and let cool...

6. Then with sterile instruments and gloves sterelized with spray, cut open along the slits and aportion seeds to agar filled mason jars.

7. Place micron filter (and then screw on mason lid?)

8. Question: what do I do for light? And humidity?

Cheers,

Andrew

 

[frogparty]

Don't fill them with water in the dishwasher, just run them normally.

How do you plan to sterilize your agar media? Pressure cooker? Autoclave? You cannot just boil agar media, it needs to be pressure sterilized!!

Sterilize your jars in autoclave/pressure cooker! Much better than microwave!!!

 

[aNisip]

Oops, yea I didn't mean to have that there... no water in the mason jar while in dishwasher...

And no autoclave but yes to pressure cooker....how should I prepare it?

And what do I do abaut the light they receive?